Re: Counterstain for Alizarin Red

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Bourassa, Patricia wrote:
> Hi, All!
> What options do I have for counterstaining with Alizarin Red?  I've gotten
> really nice deposit staining with it, but am having trouble counterstaining.

Why counterstain at all? We stain with the alizarin red about
1 minute, blot, rinse in acetone about 30 sec., then go to
about 2 changes of xylene. Presto. Done.

This is the procedure listed in the Bancroft/Stevens book.
I do this one with the students.

However, if you want a counterstain, the Dahl stain has
you stain in Alizarin red S for 30 minutes, rinse 5-6 times
in d. water, tain in light green solution 1 minute, rinse
in d.water 5-10 seconds, and run up through alcohols to xylene.

The light green solution is 1 gm light green SF yellowish
in 100 mL d. water.

This is the procedure found in Lee Luna's book. I haven't
tried it.

However, I found something interesting when comparing the
two stains. The Bancroft/Stevens book says to have the
pH at pH 4.2 to make it specific for calcium, as alizarin
red S also stains other elements. The Lee Luna book says the
alizarin red S solution should be at pH 6.5 or higher, as
slightly acid solutions will dissolve out calcium.

Guess it's a matter of sensitivity vs. specificity. And
possibly would account for the differences in staining time.

> Thanks
> -Patti Bourassa
>  Pfizer, Groton

Peggy A. Wenk, HTL (ASCP)
Anatomic Pathology
Wm. Beaumont Hospital
3601 W. 13 Mile Rd.
Royal Oak, MI 48073-6769

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